WO2003013589A1

WO2003013589A1 - Isulin-containing oral spray and the preparation method thereof

Info

Publication number: WO2003013589A1
Application number: PCT/CN2002/000342
Authority: WO
Inventors: Huibi Xu; Kaixun Huang; Qiuhua Gao; Zhonghong Gao; Jilin Yang
Original assignee: Huazhong University Of Science & Technology; Hubei Huagong Biochemistry Eng. Co., Ltd.
Priority date: 2001-08-08
Filing date: 2002-05-20
Publication date: 2003-02-20
Also published as: CN1335182A; EP1424077B1; EP1424077A4; EP1424077A1; US20040258623A1; ATE412422T1; DE60229647D1

Abstract

The invention provides an insulin-containing oral spray and the preparation method thereof. The insulin oral spray of the invention is a microemulsion which the average diameter of the particles is less than 200nm, comprising 10000u-70000u insulin, 5-50g soybean lecithin as absorption promoter, 25-80g propylene glycol as cosolvent, balancing with phosphate buffer of pH 6.8-7.8 to 1000ml emulsion. The preparation method comprises 1) adding soybean lecithin to propylene glycol, mixing sufficently, then adding phosphate buffer to the result solution, and sonicating, thus obtaining the oil phase of the emulsion; 2) dissolving insulin in phosphate buffer; 3) adding the insulin solution to the oil phase, sonicating again, thus obtaining the emulsion of the invention. The efficiency and stability of the emulsion of the invention are improved significantly, and administration of the invention is convenient.

Description

Insulin oral spray and preparation technology thereof

The invention relates to an insulin preparation and a method for producing the same, and particularly to an insulin oral spray for absorption into the oral mucosa of a human body and a preparation method thereof. Background technique

Insulin in the gastrointestinal tract is easily degraded by gastric acid and a variety of digestive enzymes. It cannot be taken orally, but can only be administered by injection. It must be injected half an hour before a meal to control blood sugar, and it must be administered for life, which makes patients feel inconvenient. Therefore, research is safe, convenient and effective, especially for non-injection preparations, which will greatly facilitate patients with disease. The non-injective route of insulin administration has become a hot topic in the pharmaceutical industry at home and abroad in the 1970s, and great progress has been made since the 1990s. There are several options for insulin administration. For example, the method of embedding an insulin pump in the peritoneum has proven to be safe and effective. It has been used in clinical practice abroad, but it is expensive and not affordable for ordinary patients. Some results have shown that the method of absorbing insulin through the bronchial mucosa has attractive application prospects, but there are still some difficulties to overcome in this method. Insulin is embedded in liposomes or polymers, and then used as an oral agent. Insulin is absorbed into the blood from the small intestinal mucosal cells to reduce blood sugar, but the bioavailability of the agent is low and the agent is easily cleared from the site of administration. Dropping is an important obstacle to the success of oral insulin. A large amount of research work is currently focused on the route of absorption and administration through the mucosa of the nose, eyes, lungs, and mouth. This work has made some progress after more than ten years of exploration, but due to the large molecular weight of insulin, the mucosa is difficult to Absorption, the clinical application of these pathways still has the problem of low bioavailability. Therefore, the selection of appropriate low-toxic and efficient absorption enhancers to increase absorption is the key to improve the bioavailability of insulin. Summary of the Invention

The invention provides an insulin oral spray and a preparation process thereof. The problem to be solved is to further improve the bioavailability of insulin through the human oral mucosa, while enhancing the stability of the preparation.

Based on the patent application of 00114318.2, the inventors further optimized the distribution ratio of each group, and adopted ultrasonic treatment for each component instead of stirring to prepare a microemulsion formulation with an average particle size of less than 200 nm.

1

Replacement page (Article 26) The insulin oral spray of the present invention is a microemulsion formulation with an average particle size of less than 200 nm. Each 1000 ml of the microemulsion of the present invention contains 1000u-70000u insulin, 5-50g of soy lecithin as a promoter, and 25-80g as a helper. The solvent is propylene glycol, and the balance is a phosphate buffer solution of pH == 6.8-7.8.

According to the insulin oral spray of the present invention, the average particle diameter of the microemulsion is preferably 100-180 nm.

The insulin oral spray of the present invention may further contain borneol-ethanol solution as a flavoring agent, and phenol as a bacteriostatic agent. That is, each 1000ml of the microemulsion of the present invention contains 15000u-60000u insulin, 20-50g of soy lecithin as a promoter, 40-80g of propylene glycol as a co-solvent, and 1.2-10g of borneol dissolved in 1-15ml of absolute ethanol. Bornol-ethanol solution of flavoring agent, 2g-5g of phenol as bacteriostatic agent, the balance is a phosphate buffer solution with pH = 6.8-7.8.

The preparation method of the insulin oral spray of the present invention comprises the following steps: taking soybean lecithin according to the formula, adding a corresponding amount of propylene glycol and stirring uniformly, and then adding a total amount of 25-60% by volume of a phosphate buffer solution to form a compound. Forming agent, ultrasonically treating the excipient for 0.5-2 hours to obtain a microemulsion oil phase; dissolve insulin in a formula proportion in a phosphate buffer solution of 40-75% by volume, and then gradually add it The microemulsion oil phase is further sonicated for 2-10 minutes.

When flavoring agents and bacteriostatic agents are added, the preparation method of the insulin oral spray of the present invention includes the following steps: taking phenol according to the formula ratio and adding it to the full amount of phosphate buffer solution; taking soybean lecithin according to the formula ratio Add the corresponding amount of propylene glycol, borneol-ethanol solution, stir and hook, and then add the total amount of 25-60% by volume of phenol-containing phosphate buffer solution to form an excipient, ultrasonically treat the excipient for 0.5-2 hours to obtain Oil phase of microemulsion; Dissolve insulin in group formula in 40-75% by volume of phenol-containing phosphate buffer solution, then gradually add it to the oil phase of the microemulsion, and continue ultrasonic treatment 2-10 minute.

In the method for preparing the insulin oral spray of the present invention, it is further characterized in that the ultrasonic treatment has an ultrasonic frequency of 18.25 KHz and an ultrasonic condition duty cycle of 30-90%.

In the preparation method of the insulin oral spray of the present invention, the temperature is controlled at 2-7 (TC) during the preparation. Insulin is a polypeptide hormone, which has poor permeability when administered directly through the oral mucosa, in order to improve its bioavailability. It is necessary to select a suitable absorption enhancer. The soybean lecithin used in the present invention is a non-toxic, safe and effective accelerator. Due to the poor water solubility of soybean lecithin, it is necessary to select a suitable co-solvent. The present invention takes the hypoglycemic effect as an indicator The solubilizing effect of propylene glycol was studied. The experimental results show that propylene glycol can increase the absorption effect of soy lecithin on oral mucosa to insulin, and their optimal ratio was determined by orthogonal experiments. The system of insulin in soybean lecithin and propylene glycol A thermodynamically stable microemulsion system is formed. Soy lecithin is also used as a carrier for surfactants and drugs in this agent. Water-soluble insulin is embedded in the lipid of soybean lecithin. Bilayer aqueous solution.

In the oral spray of the present invention, any pharmaceutically acceptable excipient may be included as long as it does not damage the properties of the microemulsion and the effect of the medicine. These pharmaceutically acceptable excipients include, but are not limited to, various diluents, solvents, emulsifiers, preservatives, stabilizers, dissolution aids, flavoring agents, fragrances, and the like.

The insulin oral spray of the present invention is non-toxic. A long-term toxicity test of oral administration of insulin oral spray in rats at doses of 4.5, 9.0, and 18.0 u-kg " ¹ , with physiological saline and excipients as the control group. After 6 months, each administration group Compared with the normal saline control group, the local and overall conditions of the animals were not significantly abnormal.

Compared with the original method, the insulin oral spray prepared by sonication has significantly improved the efficacy and stability:

1. Stability: Due to the microemulsion of insulin oral spray prepared by ultrasonic treatment, the particle size of the particles has become significantly smaller, from the original average particle size of 427.2nm to the current nano-level particles, the average particle size is less than 200nm, particle size distribution is 100-160nm. This nano-microemulsion is not only good for mucosal absorption, but also improves the stability of the preparation. From the naked eye, the insulin oral spray prepared by ultrasonic treatment was translucent and stored in a refrigerator at 2-8 ° C for 1 year. No precipitation was observed. The insulin oral spray prepared by stirring was opaque and stored in A small amount of precipitation occurred in the refrigerator at 2-8 ° C for 1 year. It can be seen that the method of ultrasonic treatment is extremely important for the preparation of microemulsions.

2. Pharmacodynamics: The present invention uses normal rabbits as a model and uses enzyme-linked immunoassay to determine the insulin content in the body. The pharmacokinetics of insulin oral sprays are calculated and their pharmacokinetic parameters are calculated. After oral mucosal administration (1.5u, kg- ¹ body weight) in rabbits, the serum insulin concentration increased rapidly, reaching a peak time of 55-70 minutes, and the peak concentration reached 145.2 ± 5.8μ / ηι1, gradually after 4 to 6 hours. Reduced to the basic level before medication, bioavailability reached 29.8%. Compared with the insulin-free excipient group, the serum insulin concentrations of the insulin oral mucosa administration group and the insulin subcutaneous injection control group were significantly different within 30 to 120 minutes (ρ <0.05). At this dose, compared with the subcutaneous injection of insulin in the control group (0.5u, kg-), the peak concentration and peak time of serum insulin were basically the same, and there was no significant difference (ρ> 0.1).

The oral spray of the present invention can be administered in a single dose or in multiple doses, preferably 3 times (every morning, middle, before dinner) or 4 times (before meals and before bedtime) every white, and sprayed 60 minutes before meals medicine. The dosage of the oral spray of the present invention is not particularly limited, and it is a conventional insulin dosage. The specific dose depends on the individual patient, bioavailability, and severity of the condition, and should be administered under the guidance of a doctor.

This medicine spray is a microemulsion, which has the characteristics of large contact area with the oral mucosa, fast absorption, good hypoglycemic effect, and safe use. This preparation is packed in a packaging bottle with a metering pump. It can convert insulin like Sprayed into the cavity like a mist, insulin adheres to the mucous membrane of the oral cavity, through which the mucosa can quickly enter the blood circulation, improve the absorption rate, and avoid the acid and enzymatic hydrolysis of the intestine and stomach, and the first-pass effect of the liver. The insulin oral spray of the present invention is a new non-injection administration method convenient for patients, which will bring convenience to the patients with diabetes, reduce the pain of injection administration, and has strong practicability. detailed description

Example 1: 1000ml insulin microemulsion

Insulin 40000U

Soy Lecithin 25.0g

Propylene glycol 75.0g

Phosphate buffer (pH = 7.4), add to 1000ml

Take soybean lecithin according to the formula, add propylene glycol, stir evenly into a thin paste, and then add 550 ml of osmate buffer solution, pH = 7.4. The above components are used as excipients in the formulation of insulin oral sprays and subjected to ultrasonic treatment. For 1 hour, the ultrasound conditions are: the duty cycle is 50%, the temperature is controlled at 40 ° C-60 ° C, and the ultrasound frequency is 20KHz. Insulin was dissolved in 350 ml of phenol-containing phosphate buffer, pH = 7.4, and slowly added to the sonicated excipient solution, and the sonication was continued for 5 minutes. As measured by a laser particle size tester, the average particle diameter was 160-180 nm.

The preparation of insulin oral spray according to this formula needs to be performed under strict aseptic conditions. The prepared spray is stored in a refrigerator at 2-8 ° C.

The insulin oral spray prepared according to the formula has a better blood glucose lowering effect. When streptozotocin-induced diabetic rats received oral doses of 1, 3, 9u * kg- ¹ , the hypoglycemic rates were 20.9%, 47.6%, and 58.8%, respectively. The oral administration doses of dioxin-induced diabetic rabbits were 0.5, 1.5, and ^ Su. Kg- ¹ , and the hypoglycemic rates were 24.9%, 52.6%, and 60.9%, respectively. Example 2: 1000mU Isulin microemulsion

Insulin 20000U

Soy Lecithin 30.0g

Propylene glycol 45.0g

Borneol 1.2g

Anhydrous ethanol 3.0ml 2.0g of phenol

Phosphate buffer (pH = 7.4), add to 1000ml

Phenol was dissolved in phosphate buffer according to the formula. Take soybean lecithin according to the formula, add propylene glycol and borneol in ethanol solution, and stir to add 555ml of phenol-containing phosphate buffer solution with pH = 7.4. After processing for 1.5 hours, the ultrasonic conditions are: the duty cycle is 60%, the temperature is controlled at 50 ° C-60 ° C, and the ultrasonic frequency is 18KHz. Insulin was dissolved in 345 ml of a phenol-containing phosphate buffer, pH = 7.4, and slowly added to the sonicated excipient solution, and the ultrasound treatment was continued for 4 minutes. As measured by a laser particle size tester, the average particle size was 160-180 nm.

The insulin oral spray prepared according to this formula was stored in a refrigerator at 2-8 ° C for one year, without precipitation.

The insulin oral spray prepared according to the formula has a better blood glucose lowering effect. The oral administration dose of streptozotocin-induced diabetic rats was 1, 3, 9u * kg- ¹ , and the hypoglycemic rates were 21.8%, 47.2%, and 56.2 respectively. When the oral dosage is 0.5, 1.5, and 4.5u ¹ , the hypoglycemic rate is 28.6%, 55.2%, and 60.7%, respectively. Example 3: 1000ml insulin microemulsion

Insulin

Soy Lecithin 25.0g

Propylene glycol 75.0g

Borneol 1.2g

Absolute ethanol 4.0ml

Phenol 2.0g

Phosphate buffer (pH = 7.0), add to 1000ml

Take soybean lecithin according to the formula, add propylene glycol and borneol in ethanol solution, and stir to add 540ml of phenol-containing phosphate buffer solution, pH = 7.0. The above components are used as excipients in insulin oral spray formulations and subjected to ultrasound. After processing for 1 hour, the ultrasonic conditions are: the duty cycle is 50%, the temperature is controlled at 40-55 ° C, and the ultrasonic frequency is 20KHz. Insulin was dissolved in 360 ml of phenol-containing phosphate buffer, pH = 7.0, and slowly added to the sonicated excipient solution, and the sonication was continued for 5 minutes. As measured by a laser particle size tester, the average particle size was 130-150 nm.

The insulin oral spray prepared according to the formula has a better blood glucose lowering effect. Induced with streptozotocin When the oral doses of diabetic rats were 1, 3, and 9 u'kg- ¹ , the hypoglycemic rates were 21.3%, 49.8%, and 60.2%, respectively. The oral administration doses of dioxin-induced diabetic rabbits were 0.5, 1.5, and 4.5u * kg- ¹ , and the hypoglycemic rates were 29.6%, 55.4%, and 62.2%. Example 4: 1000ml insulin microemulsion

Insulin 60000U

Soy Lecithin 35.5g

Propylene glycol 80.0g

Borneol 1.4g

Absolute ethanol 4.0ml

Phenol 2.0g

Phosphate buffer (pH = 7.8), add to 1000ml

Take soybean lecithin according to the formula, add propylene glycol and borneol in ethanol solution, and stir to add 545ml of phenol-containing phosphate buffer solution, pH = 7.8. The above components are used as excipients in the insulin oral spray formulation, and ultrasonicated. After processing for 2 hours, the ultrasonic conditions are: the duty cycle is 70%, the temperature is controlled at 55 ° C-70 ° C, and the ultrasonic frequency is 25KHz. Insulin was dissolved in 360 ml of phenol-containing phosphate buffer, pH = 7.8, and slowly added to the sonicated excipient solution, and the sonication was continued for 3 minutes. It was measured by a laser particle size tester, and its average particle size was 140-160 nm.

The insulin oral spray prepared according to this formula was stored in a refrigerator at 2-8 ° C for 1 year, without precipitation.

The insulin oral spray prepared according to the formula has a better blood glucose lowering effect. When streptozotocin-induced diabetic rats received oral doses of 1, 3, 9u * kg- ¹ , the hypoglycemic rates were 18.3%, 35.2%, and 44.2%, respectively. When the oral doses of dioxin-induced diabetic rabbits were 0.5, 1.5, and 4.5u * kg- ¹ , the hypoglycemic rates were 20.1%, 45.4%, and 52.2%, respectively.

Claims

Claim

1. An insulin oral spray, which is a microemulsion having an average particle size of less than 200 nm, wherein each 1000 ml of the microemulsion contains 1000u-70000U insulin, 5-50g of soy lecithin as a promoter, and 25-80g as a helper. The solvent is propylene glycol, and the balance is a phosphate buffer solution with pH = 6.8-7.8.

2. The insulin oral spray according to claim 1, characterized in that each 1000ml of the microemulsion contains 15000U-60000u insulin, 20-50g of soy lecithin as a promoter, and 40-80g of propylene glycol as a co-solvent, The balance is a phosphate buffer solution with pH = 6.8—7.8.

3. The insulin oral spray according to claim 1, wherein the microemulsion has an average particle diameter of 100-180 nm.

4. The insulin oral spray according to claim 2, characterized in that the microemulsion further contains borneol-ethanol solution as a flavoring agent, wherein the amount of borneol per 1,000 ml of the microemulsion is 1.2-10 g, anhydrous The amount of ethanol used was 1 to 15 ml.

5. The insulin oral spray according to claim 2, wherein the microemulsion further contains phenol as a bacteriostatic agent, wherein the amount of phenol in each of the microemulsions is 2 g to 5 g.

6. The method for preparing an insulin oral spray according to any one of claims 1-5, comprising the following steps:

Add 5-50 g of soybean lecithin to 25-80 g of propylene glycol, and then add a total amount of 25-60 vol% phosphate buffer solution to form an excipient. Ultrasonic treatment of the excipient for 0.5-2 hours to obtain a microemulsion Oil phase; 1000u—70,000U insulin is dissolved in 40-75 vol% phosphate buffer solution, and then gradually added to the microemulsion oil phase, and the ultrasonic treatment is continued for 2-10 minutes.

7. The preparation method according to claim 6, comprising the following steps:

Dissolve 2g-5g phenol in phosphate buffer solution;

Add 20-50g of soybean lecithin to 40-80g of propylene glycol, borneol-ethanol solution, and then add a total amount of 25-60% by volume of a phenol-containing phosphate buffer solution to form an excipient. Ultrasonicate the excipient 0.5 — 2 hours to obtain a microemulsion oil phase;

Dissolve 15000u-60000U of insulin in a phenol-containing phosphate buffer solution that accounts for 40-75% by volume of the total amount, and then gradually add it to the oil phase of the microemulsion, and continue the sonication for 2-10 minutes.

8. The preparation method according to claim 6, characterized in that the ultrasonic treatment ultrasonic frequency is 18-25 KHz, and the ultrasonic condition duty cycle is 30-90%.

9. The preparation method according to claim 7, characterized in that the ultrasonic treatment ultrasonic frequency is 18-KHz, and the ultrasonic condition duty cycle is 30-90%.

10. The preparation method according to claim 6, characterized in that the temperature during the preparation is controlled at 2 ° C.

11. The preparation method according to claim 7, characterized in that the temperature during the preparation is controlled at 2 -Ό.